Legume Genomics and Genetics 2024, Vol.15, No.3, 105-117 http://cropscipublisher.com/index.php/lgg 112 Narula, 2017; Rodriguez-Leal et al., 2017; Wan et al., 2021). The simplicity and high mutation efficiency of CRISPR/Cas9 make it an attractive option for developing new germplasm resources and enhancing agronomic traits (Arora and Narula, 2017). Additionally, advancements in CRISPR/Cas9, such as base editors and prime editors, have further improved the specificity and efficiency of gene editing, offering new opportunities for pea breeding (Biswas et al., 2021). Figure 2 CRISPR-Cas9 mediated genome editing (Adopted from Wan et al., 2021) Image caption: (I) Selection of the desired genomic DNA target, and recognition of protospacer adjacent motif (PAM) sequences before 20 bp sequences. Design of the sgRNA using online bioinformatics tools. (II) Cloning of designed sgRNAs, and binary vector construction using promoters. (III) The delivery of CRISPR-Cas editing reagents into plant cells. The vector can be transferred into the plant via Agrobacterium tumefaciens, nanoparticles, biolistic bombardment, or polyethylene glycol (PEG). Alternatively, plant RNA viruses have been used to induce heritable genome editing. When the cassette harbouring the sgRNA, RNA mobile element, and tobacco rattle virus (TRV) is transformed into the Cas9 expressing plants, the systemic spread of sgRNA will introduce heritable genome editing. (IV) Plant transformation and development of transgenic plants. (V) Genotyping of transgenic plants. (VI) Transgene-free plants with the desired mutation are obtained (Adopted from Wan et al., 2021) 8.3 Case studies of successful breeding programs Several successful breeding programs have demonstrated the potential of modern breeding techniques in improving pea varieties. For example, a study on the genetic diversity and trait genomic prediction in a pea diversity panel utilized newly developed SNP markers to characterize diverse pea accessions and predict phenotypes such as TSW, NSeed, and BegFlo. The study showed that high-throughput SNP arrays could significantly enhance the efficiency of marker-assisted selection in peas (Burstin et al., 2015). Another case study highlighted the application of CRISPR/Cas9 in crop quality improvement, where the technology was used to modulate traits such as appearance, palatability, and nutritional components in various crops (Liu et al., 2021). This approach has accelerated crop breeding progress by enabling precise gene editing and the development of non-genetically modified (Non-GMO) crops with desired traits (Jaganathan et al., 2018). Overall, the integration of genomic selection and CRISPR/Cas9 technology in pea breeding programs has shown great promise in enhancing yield potential, improving stress resistance, and developing high-quality pea varieties. These advancements underscore the importance of modern breeding techniques in addressing the challenges posed by global climate change and meeting the demands of sustainable agriculture.
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